[Histonet] need tips for cross-sectioning of cortical bone

[Joseph Saby]

Brett-

Most wrinkles in decalcified bone sections come from stretching of the decalified bone that occurs during the sectioning process.  I would suggest a rather simple solution.  Allowing the sections to flatten on the waterbath might take longer or a higher temperature.  If paraffin surrounding the bone section seems to be containing it, not allowing it to expand to eliminate the wrinkles, gently tease it off.  After all, you want the bone section, not the paraffin.  A room temperature water bath (or 30% EtOH) to lay out the sections on to tease out any wrinkles before transfering the sections to the warm waterbath may also help.

I hope this helps!

Joe Saby, BA HT




________________________________
From: Adam . <anonwums1 <@t> gmail.com>
To: "Connolly, Brett M" <brett_connolly <@t> merck.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Sent: Thu, April 22, 2010 11:12:26 AM
Subject: Re: [Histonet] need tips for cross-sectioning of cortical bone

Cutting bone is very hard, and I'm by no means an expert at it. Assuming the
blocks are properly fixed and decalcified, the best thing I've found is to
put the blocks at -20C for 5-10 mins to cool them, then right before you cut
them, rub a little ice water on the face of the block. That should help you
get some nice clean cuts. If the sections become hard to cut again, reapply
the ice water. If that stops working, back in the freezer they go.

Adam

On Thu, Apr 22, 2010 at 9:58 AM, Connolly, Brett M <brett_connolly <@t> merck.com
> wrote:

> A colleague is having trouble getting wrinkle-free sections of
> decalcified, paraffin embedded femur.
>
> Any tips??
>
> Thanks,
>
> Brett M. Connolly, Ph.D.
> Molecular Imaging Team Leader
> Merck & Co., Inc.
> PO Box 4, WP-44K
> West Point, PA 19486
> tel. 215-652-2501 fax. 215-993-6803
> brett_connolly <@t> merck.com
>
>
>
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